Cryopreservation of canine testicular tissue: from fragmentation to sample processing

Abstract

The deep bond between humans and dogs drives the pursuit of innovative methods to preserve valuable traits across different breeds. Among these techniques, testicular tissue cryopreservation emerges as a promising alternative to support canine reproduction, especially when traditional semen collection methods are impractical. This strategy not only enables the unlimited generation of male gametes but also facilitates the establishment of germplasm banks, which are crucial for both animals of significant genetic value and the conservation of endangered species. This study focuses on refining the cryopreservation of testicular tissue in dogs, a critical step in ensuring the fertility and genetic diversity of these animals. The process involves meticulous dissection of the testicles, followed by tissue segmentation and exposure to slow freezing or vitrification methods, using cryoprotectants to minimize cellular damage. Specifically, solid surface vitrification (SSV) is highlighted for its efficiency and lower cost. After cryopreservation, the tissue is carefully reheated, with strict control over time and temperature to prevent cellular damage. The analysis of viability, integrity, and metabolic functionality of the tissues is conducted using advanced techniques, such as laser confocal microscopy and histomorphological evaluations. Contributing significantly to advancements in this research area, this study provides crucial data for grounding future work and the practical application of these findings in clinical contexts, opening new perspectives for canine reproduction and the conservation of endangered species.